Thursday, April 25, 2013

Applications Thursday: Simultaneous dual emission detection of luciferase reporter assays

Schematic of a luciferase reporter assay

Dual luciferase reporters have become the go to assay to assess changes in gene expression that result from altering the activity of components of a biological pathway. The standard approach employs sequential detection of one luciferase that relates to the studied subject and a second luciferase that is used to indicate cell viability and transfection efficiency.

In BMG LABTECH application note 233, we describe the use of the POLARstar Omega to perform the Thermo Scientific Pierce Luciferase Dual-Spectral Assays. Since these assays employ spectrally resolved pairs of luciferase enzymes it allows the user to perform one-step detection of both enzymes. The capability to perform simultaneous dual emission detection allows for one read assessment using the POLARstar Omega.

Gaussia-Firefly Dual-Spectral
Luciferase Assay
Selecting filter pairs that allowed for sensitive detection of each luciferase pair while limiting the interference between them was key to the success of this application. Application note 233 clearly describes the final filter sets employed that allow for the accurate measurement of each reporter over a 100,000-fold concentration range for three possible combinations of spectrally resolved luciferases.

To find out more about the capabilities of the POLARstar Omega please visit